hIL21/hIL21R

Fig.1 Detection of IL21 expression in splenocyte by RT-PCR. 

Wild type: only one band at 272 bp with primers F1/R1 (mIl21);

Homozygous: only one band at 304 bp with primers F2/R2 (hIL21); 

Abbr. M, DNA marker; HO, homozygous; WT, wild type.

Fig.1 Detection of human IL21 (A) and mouse IL21(B) expression in splenocyte culture supernatants of WT C57BL/6 mice and HO hIL-21/hIL-21R mice by ELISA.  Splenocytes were isolated from male 6-week-old wild-type (WT) and HO/HO hIL-21/hIL-21R mice, then seeded in 12-well plates at a density of 1×106 cells/well in 1 mL culture medium. Following in vitro stimulation with 1 μg/mL CD3 antibody and 20 ng/mL anti-mouse IL6 antibody for 72 h,  IL21 levels in the culture supernatants were measured using anti-hIL21 ELISA kit and anti-mIL21 ELISA kit.

Abbr.  HO, homozygous; WT, wild type; N.D. not detected.

Fig.3 Detection of IL-21R expression on T cells and B cells of  of WT C57BL/6 mice and HO hIL21/hIL21R mice by FACS. Spleenocytes were collected from wild-type C57BL/6 mice (female,11 weeks old) and homozygous hIL21/hIL21R humanized mice (female,11 weeks old), and analyzed by flow cytometry with anti-mIL-21R and anti-hIL-21R.

Fig.4 Detection of IL-21R expression on T cells and B cells of  of WT C57BL/6 mice and HO hIL21/hIL21R mice by FACS. Spleenocytes were collected from wild-type C57BL/6 mice (female,11 weeks old) and homozygous hIL21/hIL21R humanized mice (female,11 weeks old), stimulated with 1μg/ml anti-CD3 and 1μg/ml anti-CD28 for 48 h in vitro and analyzed by flow cytometry with anti-mIL-21R and anti-hIL-21R.

Fig.5 Human IL-21 induced the phosphorylation of Stat3 in CD3+ T cells, CD4+ T cells and CD8+ T cells in the C57BL/6 WT mouse (11 weeks old) and hIL21/hIL21R knockin mice (11 weeks old). Splenocytes were collected from the spleen, treated with different concentrations of Human IL-21 for 15 min and analyzed by flow cytometry.